Comparative Study on Biologic and Immunologic Characteristics of the Pancreas Islet Cell between 24degrees C and 37degrees C Culture in the Rat.
- Author:
Song Cheol KIM
1
;
Duck Jong HAN
;
Ik Hee KIM
;
Kyung Ok WOO
;
You Me WE
;
Seo Young KANG
;
Ji Hae BACK
;
Yang Hee KIM
;
Dong Gyun LIM
Author Information
1. Department of Surgery, Ulsan University College of Medicine & Asan Medical Center, Seoul, Korea.
- Publication Type:Comparative Study ; Original Article
- Keywords:
Diabetes mellitus;
Pancreas islet cell transplantation;
Low temperature culture
- MeSH:
Animals;
Annexin A5;
Apoptosis;
Cell Death;
Cell Proliferation;
Cell Survival;
Diabetes Mellitus;
Endocrine Cells;
Glucose;
Humans;
Insulin;
Interleukin-10;
Interleukin-4;
Islets of Langerhans Transplantation;
Islets of Langerhans*;
Lymphocytes;
Pancreas*;
Rats*;
Tumor Necrosis Factor-alpha
- From:The Journal of the Korean Society for Transplantation
2004;18(2):125-133
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: Islet cell transplantation, as an alternative approach to endocrine cell replacement to treat the diabetes mellitus, has received significant attention because it holds several advantages over whole gland transplantation. However cell damage from islet isolation and immunologic rejection after transplantation prevent from successful clinical application for diabetic patients. Culture of cells at low temperature has known to stabilize the cell viability, and to decrease the immunologic antigenicity. Aim of this study is to investigate the effect of culture at 24oC on cell viability, cellular function, immunogenicity and cytokine profiles in rat pancreas islet. METHODS: Pancreas islets were isolated from Lewis rat and cultured at 24oC or 37oC during 14 days. Islet recovery after culture period was counted as islet equivalent number, and islet viability was examined with fluorescent vital staining (FDA/PI). Islet function was measured with glucose stimulation test. Annexin V expression and MHC class I and II expression were measured with flow cytometric assay for apoptosis and immunogenicity respectively. Lymphocyte cell proliferation through mixed lymphocyte islet culture was examined with WST-1 proliferation assay. Cytokine profiles were analyzed with quantitative real time RT-PCR. All these parameters were measured on 1, 3, 5, 7, 14 culture days after islet isolation. RESULTS: Islet recovery was higher in islet cultured at 24oC than in islet cultured at 37oC without change of viability. Insulin secretion after glucose stimulation was more effective in 24oC culture condition. Decrease of apoptotic cell death was demonstrated in 24oC cultured islet. MHC class I and II expression on islets and lymphocyte proliferation when cocultured with islets were less prominent in 24oC cultured islet. TNF-alpha and IL-4 cytokine expression was higher in islet cultured at 24oC than in islet cultured at 37oC. IL-1beta and IL-10 cytokine expression were similar in both culture condition. CONCLUSION: This study demonstrated that cell recovery and function are increased in islet cultured at 24oC than in islet cultured at 37oC while antigenicity and proinflammatory cytokine expression are decreased. Low temperature culture can be a good approach to prevent the loss of islet mass, and to reduce the immunologic rejection of transplanted islet for successful clinical islet transplantation.
